№ files_lp_3_process_9_63684
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Laboratory protocol detailing reagents, preparation of RPPA working solution, cell pellet lysis, protein quantification, SDS sample buffer addition, denaturation, labeling, storage, and submission procedures for Reverse Phase Protein Array analysis.
Title: Protocol for Lysis of Cell Pellets for Reverse Phase Protein Array (RPPA)
Language: English
Document Type: Laboratory protocol
Technique: Reverse Phase Protein Array (RPPA)
Application: Cell pellet lysis and sample preparation for protein analysis
Reagents Provided: RPPA Lysis Buffer; 5X Protease Inhibitors; 5X Phosphatase Inhibitors; 2X SDS Sample Buffer; 4X SDS Sample Buffer
Reagents Required but Not Supplied: β-mercaptoethanol; 1.5 ml microcentrifuge tubes
Storage Conditions: 4°C; -20°C; -80°C
Assays Referenced: BCA assay; Bradford assay
Required Equipment: Microcentrifuge; vortex; rotator; heating block or thermocycler
Target Protein Concentration: 1.1–3 mg/ml for lysates; 0.5 mg/ml final concentration for RPPA samples
Sample Type: Cell culture pellets
Submission Requirement: RPPA Sample Submission Form via email prior to delivery
Delivery Conditions: Samples delivered on dry ice to the Core facility
Price: 8 / 10 USD
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Supplemental Methods for Cell Lysis, Transfections, and Protein Analysis in Mouse Tissues
Year: 2023
Organism: Mus musculus
Tissue types: Optic nerve, cerebellum, C2C12 cells
Techniques: Cell lysis, transfection, RNA extraction, western blotting, immunofluorescence, RT-PCR
Reagents and tools: RSB-100 buffer, TriZOL, Turbo DNase, Lipofectamine2000, plasmids (pcDNA3.1, tdTomato, Qk reporters), siRNAs, morpholinos, primary and secondary antibodies
Institution: University of California, Santa Cruz
Animal model: Adult wild-type 129-C57BL/6 mice
Ethics approval: UCSC Institutional Animal Care and Use Committee
Target proteins: QKI isoforms (QKI-5, QKI-6, QKI-7), GAPDH, Myc, HA, histone H3, tdTomato
Assays: Dual Glo Luciferase Assay, BioAnalyzer quantitation, RT-qPCR
Method type: Experimental protocol
Supplementary information: Details on plasmid construction, antibody sources, and subcellular fractionation
Lysis, or Friendship
Author: Plato
Translator: Benjamin Jowett
Literary Form: Philosophical dialogue
Genre: Classical Greek philosophy
Main Characters: Socrates; Menexenus; Hippothales; Lysis; Ctesippus
Narrator: Socrates
Setting: Newly erected palaestra outside the walls of Athens
Place of Action: Athens, ancient Greece
Historical Context: Classical Greek period
Philosophical Theme: Nature of friendship and affection
Original Language of Work: Ancient Greek
Text Source Form: English translation of a classical philosophical dialogue
ONR Project Assessment Report PR-01156 Fissile Validation for Transport of Unirradiated Pellets of Uranium Oxides (Package Model GP-01) - Transport Package Design Approval
Year: 2023
Region / City: United Kingdom
Subject: Transport package design approval
Document Type: Assessment Report
Author: Office for Nuclear Regulation (ONR)
Target Audience: Regulatory authorities, Nuclear transport agencies
Period of Validity: Until 30 November 2028
Approval Date: 25 February 2023
Amendment Date: 14 December 2023
Topics: RPPA Lysis Pellets